Column and Thin Layer Chromatography: the Separation of Spinach Pigment

Column and Thin Layer Chromatography: The Separation of Spinach Pigment

AbstractSpinach extract was separated into fractions containing compounds of similar polarity by column chromatography. Based on solid-liquid phase partitioning, this separation technique exploited the different polarity of the compounds in the spinach extract. Three fractions with different colors were obtained. The extract and its fractions were analyzed using thin layer chromatography (TLC). The TLC results showed that there was one compound (Rf=0.979) in the first fraction; there were three compounds (Rf1=0.839, Rf2= 0.691, Rf3=0.149) in the second fraction; there was one compound (Rf=0.017) in the third fraction. The separations of compounds which went to the first and third fraction were relatively satisfying, while the second fraction had several kinds of compounds. Introduction

Chromatography is the separation of compounds or ions by distribution between two phases—a mobile phase and a stationary phase. The technique is based on the differential absorptivities of the constituents between these two phases, due to different properties of the compounds to be separated and the nature of the two phases involved. If one constituent adheres more to the stationary phase than the mobile one, separation will be achieved.

There are several different types of chromatography, such as thin layer chromatography (TLC), gas chromatography (GC) and column chromatography. All the chromatographic methods are based on partitioning of molecules between a stationary phase and a mobile phase. In order to measure the amount of partitioning between the two phases, each compound has an unique Partition Coefficient (Kp), which is defined as the ratio of concentrations of the compound between the stationary phase and the mobile phase: Kp=[x]sp/[x]mp. Therefore, if Kp is greater than 1, the substance adheres more to the stationary phase; if Kp is smaller than 1, the substance adheres...

You May Also Find These Documents Helpful

...Laboratory 1
Column and ThinLayerChromatography: The Separation of SpinachPigments
Russ Hoburg
02/15/2012
Abstract
The main goal in doing the column and thinlayerchromatography was to separate spinach extract into its components based on polarity and then to analyze the components. The separation of the spinach extract was done using the columnchromatography with the wet/slurry packing method. Alumina was inserted into the column to act as the stationary phase. After the components were separated and the solvents boiled out, the thinlayerchromatography was performed to analyze the components of the spinach extract. This was done using a TLC plate and spotting each of the components on to the plate as well as spotting the standard spinach extract. A beaker was used with the developing solvent, filter paper, and TLC plate inside and a watch glass over top of the beaker to perform the thinlayerchromatography. After the solvent front (30% ethyl acetate in hexane) reached about an inch from the top, the TLC plate was removed from the beaker and the developing spots were marked. After implementing the...

...﻿
The Separation of SpinachPigments via Column and Thin-LayerChromatography
5/29/14
Abstract:
Spinach extract was separated into fractions containing compounds of similar polarity via columnchromatography. Thinlayerchromatography was then used to analyze the extract and its separated components. Experimental results showed that the retention factor for carotene (0. 0.86) was the largest, followed by xanthophyll (0.38) and then chlorophyll (0.14) being the smallest.
Introduction:
Chromatography exploits differences in the physical properties, such as boiling point and/or polarity, of components in a mixture and separates the different compounds or ions by distribution between two phases, a stationary and a mobile phase (Padias 162). The methodology behind chromatography is based on the variant absorbability of the constituents between these two phases, due to different properties of the compounds to be separated and the nature of the two phases involved. The stationary phase is constantly washed with a mobile phase, and over time the added mixture will separate into its substituent components based on their physical properties, or whether a particular component adheres better to the mobile phase or the stationary...

...Laboratory - CH 201 (2010-2011) Experiment 4B
Column and Thin-LayerChromatography of Capsicum Frutescens L. Pigments
John Cyril Abanto*, Vernalyn Abarintos and Clarice Gail Abella
Department of Chemistry, College of Science
University of Santo Tomas, Espana Street, Manila 1050
Date Submitted:
September, 2010
____________________________________________________________
_________________________________
Abstract: The experiment was done to separate and analyze the components of chili pepper. Chromatography was used because of its powerful technique in separating mixtures. In this experiment the Chili pepper pigments was extracted using DCM, the extract was then introduced into the column and eluate was collected, a technique called columnChromatography. Using the ThinLayerChromatography the purity of the components was tested. The UV lamp was used to spot the UV visible components and the Retention Factor was computed.
Keywords: ColumnChromatography, Thin-LayerChromatography, Retention Factor ____________________________________________________________
_________________________________
1. Introduction
Chromatography (color writing) is a separation technique...

...COLUMN AND THINLAYERCHROMATOGRAPHY
Group 9
RAMOS, Sharina Joy; REYES Aina Marie; REYES Jallisa Maan;
RUBIO, John Michael; SABINO Patricia Anne; SANTOS, Carlos Rafael
ABSTRACT
To separate the colored components of siling labuyo and to determine the purity of the components, the students performed a column and thin or solid-liquid chromatography procedure. The solid may be almost any material that does not dissolve in liquid phase. But for this experiment, the solid used by the students was silica gel. Chromatography is a method of separating mixtures and at the same time useful in the identification of compounds. It is based on both solubility and absorptivity. The students were able to extract, more or less, six (6) colors and was further broken down into three (3) colors specifically yellow, red and orange. Using a precoated TLC plate, the students then was able to observe and measure the Rf values of the different components of the siling labuyo through the use of the TLC plates with eluates applied and was then placed in the developing chamber. At the end of the experiment, the students were able to discover the different components of the siling labuyo, its purity and the Rf values of each components of the red siling labuyo.
INTRODUCTION
The experiment was done to separate the colored components of siling labuyo using column...

...﻿Column and ThinLayerChromatography
Beverly
Abstract:
Plant pigments were separated and concentrated from a crude spinach extract through the use of columnchromatography and an eluatropic series of hexanes, hexane/acetone, and methanol. The pigments were analyzed using thinlayerchromatography with a 30% ethyl acetate/hexane developing solvent.
Introduction:
Chromatography is a technique used to separate a mixture of two or more components based on differences in their physical properties. It can be used as a method of purification (and can be used on a small scale) along with analyzation of mixtures. Chromatography exploits differences in the physical properties, such as boiling point and/or polarity, of components of a mixture in order to separate them between the mobile and stationary phases. A stationary phase is constantly washed with a mobile phase, and over time the added mixture will separate into its substituent components based on their physical properties, or whether a particular component adheres better to the mobile phase or the stationary phase.
Components of a mixture partition between the two phases nu explanation of the partition coefficient relating to chromatography, where the coefficient of...

...Introduction
Chromatography is a technique used to separate a mixture into its individual components. In this experiment, chromatography is done by using a thinlayerchromatography (TLC) plate and filter paper. The word ‘chromatography’ itself means the separation of colours. However, nowadays, both coloured and colourless compound are used in chromatography by various methods.
In chromatography, the separation process of mixture is mainly based upon the polarity and molecular weight of sample. Then, the separation involves mobile phase and stationary phase. The mixture to be separated is applied to an immovable solid surface, which is the stationary phase, such as TLC plate and the filter paper. Then, suitable solvent is then added to ‘wash’ the porous solid by the flow of solvent, which is the mobile phase. The mobile phase can be liquid (in liquid chromatography) which is in this experiment, or gas (as in gas chromatography).
The porous solid, which is the stationary phase, adsorbs different substances depending on the polarity of the substance adsorbed to it. The term “adsorption” means the adhesion or stickyness of a substance to the surface of another substance. As mentioned, chromatography is based on polarity. So, a mixture of compound is separated or adsorbed...

...ThinLayerChromatography of the Unknown Analgesic
Jessica Bajao*, Phoebe Abalos, Kevin Antiga, Carmelus Aseneta
3-Biology 2
College of Science, University of Santo Tomas, Manila, Philippines
Abstract
The group used five different analgesics in this experiment: aspirin, acetaminophen, ibuprofen, caffeine, and mefenamic acid. Six analgesics were spotted on the TLC plate including the unknown. After the development of the TLC plate, it was placed under ultraviolet light for the spots resulted to be traced. The distance from the origin to the solvent front and the distances from the origin to the center of each spot formed were measured to calculate for the retention factor. The resulted retention factor of the unknown is 0.71 which is similar to ibuprofen which also has a retention factor of 0.71. Using thinlayerchromatography, we identified the unknown analgesic to be ibuprofen.
Introduction
[9] The history of chromatography begins during the mid-19th century. Chromatography literally meant color writing which was primarily used for the separation of plant pigments such as xanthopyll. New types of chromatography were developed during the 19th century (and before) the first true chromatography is attributed to the Russian botanist, Mikhail Semyonovich Tsvet. He separated plant...

...Experiment 5
Title : ThinLayerChromatography
Objectives:
i. To distinguish polar and non-polar solvents.
ii. To familiar with the analysis technique by using the thinlayerchromatography.
iii. To differentiate the retention factor, Rf for different compounds.
[pic]
Result:
|Compound |Distance traveled by the compound |
|o-nitroanaline |2.45 |
|p-nitroaniline |1.70 |
|Unknown sample |2.45 |
Calculation:
The Rf value for each dye is then worked out using the formula:
[pic]
o-nitroanaline:
Rf = 2.45
3.00
= 0.82
p-nitroaniline:
Rf = 1.75
3.00
= 0.58
Unknown sample:
Rf = 2.45
3.00
= 0.82
Discussion:
Silica gel is a form of silicon dioxide (silica). The silicon atoms are joined via oxygen atoms in a giant covalent structure. However, at the surface of the silica gel, the silicon atoms are attached to -OH groups.
So, at the surface of the silica gel you have Si-O-H bonds instead of Si-O-Si bonds. The diagram shows...